Search PubMed⌕ Search

Biomedical subjects

J Santos

Publications and source records attributed to J Santos.

At least 73 records · Page 4Linked to original sources
PubMed1999

Loss of heterozygosity at the proximal-mid part of mouse chromosome 4 defines two novel tumor suppressor gene loci in T-cell lymphomas.

Recent studies in our laboratory reported frequent loss of heterozygosity (LOH) on mouse chromosome 4 in T-cell lymphomas, identifying three candidate tumor suppressor regions (TLSR1-3). To determine the possible existence of other tumor suppressor gene loci on the proximal-mid part of chromosome 4 and to clarify whether the p16(INK4a) (alpha and beta) and p15(INK4b) genes are the inactivation targets of deletion at TLSR1, we have tested 73 gamma-radiation-induced T-cell lymphomas of F1 hybrid mice by LOH analysis. Frequent LOH was found at the INK4a and INK4b loci and the surrounding markers D4Mit77, D4Mit245 and D4Wsm1. In addition, we identified two distinct regions of significant allelic losses in the proximal-mid part of chromosome 4, defined by the markers D4Mit116 (TLSR4) and D4Mit21 (TLSR5). Taken together, this evidence and our previous data indicate the existence of at least five different candidate sites for tumor suppressor genes on chromosome 4, thus revealing a main role for this chromosome in the development of mouse T-cell lymphomas.

Alleles↗
PubMed1999

Mouse p73 gene maps to the distal part of chromosome 4 and might be involved in the progression of gamma-radiation-induced T-cell lymphomas.

We have isolated and sequenced a DNA fragment of about 12 kb that comprises exons 5-14 of the mouse p73 gene. We have identified four polymorphic markers, and one of them has been used to genetically map p73 to the distal part of chromosome 4. Previously, we have reported that gamma-radiation-induced T-cell lymphomas undergo frequent loss of heterozygosity around marker D4Mit205b at the distal part of chromosome 4. Based on this, we have performed loss of heterozygosity analysis in a set of T-cell lymphomas, and we have found allelic losses of p73 in 32.6% (16 of 49) of the tumors analyzed. Interestingly, allelic losses occur concurrently at both p73 and D4Mit205b, thus suggesting that p73 could be specifically inactivated in radiation-induced T-cell lymphomas.

Alleles↗
PubMed1999

Hypermethylation of the cell cycle inhibitor p15INK4b 3'-untranslated region interferes with its transcriptional regulation in primary lymphomas.

The cyclin-dependent kinase inhibitor p15INK4b has been shown to be involved in human and rodent tumors and seems to act as a tumor suppressor gene in hematological malignancies. Alterations of this gene in tumors include mainly homozygous deletions and hypermethylation of the CpG island in the promoter region. In this work, we describe a new area sensitive to methylation in the 3' untranslated region (UTR) of the murine p15INK4b gene. This region shows different levels of methylation depending on the tissues, being relatively highly methylated in brain and gut, and weakly methylated in liver, spleen or thymus. DNA methylation and expression is similar in both maternal and paternal alleles indicating no imprinting effect. Although methylation of the p15INK4b 3'-UTR is low in normal thymus, increased levels (up to 100%) of specific methylation in this region are found in up to 30% of radiation- or carcinogen-induced thymic lymphomas, correlating with decreased gene expression. Hypermethylation of the p15INK4b 3'-UTR frequently occurs in tumors with loss of heterozygosity (LOH) but without methylation of the promoter CpG island or intragenic mutations. Furthermore, in vitro CpG methylation of the 3'-UTR produces reduced levels of a luciferase reporter in cultured cells. Methylation of two CpG sites in a 120 bp region is sufficient to interfere with transcription of the reporter gene. These data suggest that although the levels of p15INK4b in normal tissues can be mainly determined by promoter regulatory elements, strong hypermethylation of the 3'-UTR can interfere with transcription. Thus, hypermethylation of the 3'-UTR may explain the lack of p15INK4b gene expression in a subset of tumors with no promoter methylation and could be a new alternative mechanism for tumor suppressor gene inactivation in tumorigenesis.

3' Untranslated Regions↗
PubMed1999

Analysis of the INK4a/ARF locus in non-Hodgkin's lymphomas using two new internal microsatellite markers.

Inactivation of the INK4a/ARF locus is a frequent event in non-Hodgkin's lymphomas (NHLs), which may be attributed to deletion, point mutation, and 5' CpG methylation at its promoter region. In the present study we evaluated the occurrence of deletions and genetic instability of INK4a/ARF locus in 30 paired normal and tumor samples of B cell NHLs by conducting an allelotypic analysis with two new polymorphic markers, one located at the intron 1 of p16INK4a gene and the other one placed downstream exon 1beta of p19ARF. Comparison of these results with those obtained in a previous paper using flanking markers (D9S171, D9S942, D9S958 and IFNA) allowed us to detect two new cases of microsatellite instability (L-446 and L-442), and to confirm the occurrence of LOH at the INK4a/ARF locus in one tumor (M-3770). On the contrary, this locus is not affected in three different tumors (L-421, L-272 and L-159) which exhibited LOH at some of the flanking markers.

Chromosome Mapping↗
PubMed1999

Cooperative alterations of Rb pathway regulators in mouse primary T cell lymphomas.

Alterations in the Rb pathway have been described in many different tumors. In order to study this cell cycle regulatory mechanism in murine T cell lymphomas, we have analyzed the RNA and protein expression of the cyclin D1, cdk4 and retinoblastoma genes in primary tumor samples. We have detected overexpression of the cyclin D1 gene and deficient expression of the retinoblastoma gene in 42 and 28% of these tumors, respectively. The immunohistochemical analysis showed that these RT-PCR results are correlated with a significant increase in the number of positive cells for cyclin D1 and a moderate decrease in the expression of Rb protein, respectively. The analysis of cyclin D1, Rb, p15(INK4b) and p16(INK4a) showed that 75% of lymphomas had alterations in these genes and indicates that the Rb pathway is frequently altered in mouse primary T cell lymphomas. Moreover, 31% of lymphomas presented simultaneous alterations in at least two of these genes, suggesting the importance of concurrent alteration of different Rb pathway regulators. In addition, we have characterized these samples for mutational status of the N-ras and K-ras genes. We have only detected mutations in codon 12 of K-ras in six of 49 lymphomas (12%). Interestingly, five of these lymphomas also showed alterations in at least one of the Rb pathway regulators analyzed here. Taken together, these data suggest that deregulation of the Rb pathway regulators and/or oncogenic activation of K-ras may represent a common important clue in progression of murine T cell lymphomas.

Animals↗
PubMed1999

Predictive capacity of carbamazepine pharmacokinetic parameters in a Portuguese outpatient population.

The individualization of anticonvulsant therapy regimens can contribute to the implementation of appropriate carbamazepine (CBZ) maintenance doses in epileptic patients. An accurate method for the prediction of concentrations based on a determination of parameters and serum concentrations could be of clinical relevance in the management of epilepsy. In this study, we retrospectively evaluated the predictive performance in an adult outpatient population of six different methods, representing six sets of CBZ pharmacokinetic parameters selected according to the literature using a Bayesian computer program (PKS System; Abbott Laboratories, Abbott Park, IL, USA). The study involved 50 patients with two or more available concentrations selected under several inclusion criteria. The patients were taking CBZ (between 200 and 1600 mg/d) in monotherapy or polytherapy regimens and had no hepatic or renal disease. Steady state concentrations were predicted according to the use of prior information and using one and two feedback patient concentrations. Accuracy and precision were assessed by mean prediction error (ME), mean squared prediction error (MSE) and root mean squared prediction error (RMSE). The analysis showed CL = 0.067 L/hour/kg and Vd = 1.19 L/kg as the most accurate and precise set of pharmacokinetic parameters, presenting the highest percentage of clinically acceptable estimates (error < 2 microg/mL). Additionally, predictions based on one measured feedback concentration were found to be more accurate and precise than prior population-based predictions; the use of two previous patient concentrations further improved predictive capacity but failed to show a significant difference when compared with predictions based on one measured feedback concentration. In conclusion, the adoption of the previously mentioned set of parameters as population estimates and the use of at least one feedback concentration through the Bayesian approach seems to be essential for a better CBZ use in clinical practice. Finally, despite the obtained results, we believe that the Portuguese pharmacokinetic parameter determination of antiepileptics should be carried out to improve the rationale and cost-effectiveness of anticonvulsant therapy.

Adolescent↗
PubMed1999

Characterisation of immune mediator release during the immediate response to segmental mucosal challenge in the jejunum of patients with food allergy.

BACKGROUND: Food allergy is a common complaint among patients with a broad spectrum of abdominal and extra-abdominal symptoms that must be distinguished from other more common non-immunological food intolerances. AIMS: To investigate whether human intestinal hypersensitivity reactions are associated with detectable release of inflammatory mediators from activated cells, which may serve as a biological marker of true allergic reactions. PATIENTS/METHODS: In eight patients with food allergy and seven healthy volunteers, a closed-segment perfusion technique was used to investigate the effects of jejunal food challenge on luminal release of tryptase, histamine, prostaglandin D(2), eosinophil cationic protein, peroxidase activity, and water flux. RESULTS: Intraluminal administration of food antigens induced a rapid increase in intestinal release of tryptase, histamine, prostaglandin D(2), and peroxidase activity (p<0.05 v basal period) but not eosinophil cationic protein. The increased release of these mediators was associated with a notable water secretory response. CONCLUSIONS: These results suggest that human intestinal hypersensitivity reactions are characterised by prompt activation of mast cells and other immune cells, with notable and immediate secretion of water and inflammatory mediators into the intestinal lumen. Analysis of the profile of markers released into the jejunum after food provocation may be useful for the objective diagnosis of food allergy.

Adult↗
PubMed1999

Corticotropin-releasing hormone mimics stress-induced colonic epithelial pathophysiology in the rat.

We examined the effect of stress on colonic epithelial physiology, the role of corticotropin-releasing hormone (CRH), and the pathways involved. Rats were restrained or injected intraperitoneally with CRH or saline. Colonic segments were mounted in Ussing chambers, in which ion secretion and permeability (conductance and probe fluxes) were measured. To test the pathways involved in CRH-induced changes, rats were pretreated with hexamethonium, atropine, bretylium, doxantrazole, alpha-helical CRH-(9-41) (all intraperitoneally), or aminoglutethimide (subcutaneously). Restraint stress increased colonic ion secretion and permeability to ions, the bacterial peptide FMLP, and horseradish peroxidase (HRP). These changes were prevented by alpha-helical CRH-(9-41) and mimicked by CRH (50 microgram/kg). CRH-induced changes in ion secretion were abolished by alpha-helical CRH-(9-41), hexamethonium, atropine, or doxantrazole. CRH-stimulated conductance was significantly inhibited by alpha-helical CRH-(9-41), hexamethonium, bretylium, or doxantrazole. CRH-induced enhancement of HRP flux was significantly reduced by all drugs but aminoglutethimide. Peripheral CRH reproduced stress-induced colonic epithelial pathophysiology via cholinergic and adrenergic nerves and mast cells. Modulation of stress responses may be relevant to the management of colonic disorders.

Animals↗
PubMed1999

Treatment of morbid obesity with adjustable gastric band: preliminary report.

BACKGROUND: The goal of surgery for morbid obesity is to achieve a good and durable loss of weight and improve health. Previous studies have demonstrated a significant weight loss for the Swedish adjustable gastric band (SAGB). PATIENTS AND METHODS: Between November 1996 and April 1998, 18 morbidly obese patients underwent SAGB laparoscopically. Their mean age at surgery was 35 years. The mean preoperative weight was 128 kg (range 89-163), and the mean body mass index was 50.4+/-9. Comorbidity was present in 13 patients. RESULTS: One gastric perforation occurred, and in one patient it was not possible to create the pneumoperitoneum. Regarding late morbidity, one intragastric migration and one slippage of the band occurred. There was no mortality. CONCLUSION: The low morbidity, the good results with weight loss, and the improvement in comorbidity lead the authors to believe that Swedish adjustable gastric banding for the treatment of morbidly obese patients is a successful means of losing weight and improving general health.

Adult↗
PubMed1999

Effects of bovine somatotropin and evaporative cooling plus shade on lactation performance of cows during summer heat stress.

Thirty-two Holstein cows (8 per treatment) averaging 195 d in milk were assigned to 70 d of treatment on the basis of production during a 14-d pretreatment period, which was used for covariate analysis. The experiment was a randomized block design with a 2 x 2 factorial arrangement of treatments. Factors were normal shade or shade plus evaporative cooling with pressurized spray, plus with or without the administration of bovine somatotropin (bST). Cows receiving bST were injected with 500 mg of bST every 14 d. All cows were fed the same total mixed rations twice daily at approximately 10% in excess of appetite, and water was offered free choice. There were no interactions between bST and the cooling system for any of the variables measured. Milk yield was increased by bST and tended to be greater for cooled cows. Fat percentages were increased by bST, and yields of fat, protein, and 3.5% fat-corrected milk, and the efficiency of conversion of dry matter to milk, whereas evaporative cooling increased body weights and protein yields, but decreased SNF and milk protein percentages. Rectal temperatures and respiration rates also were lower for cooled cows. And, bST increased nonesterified fatty acids in blood serum, suggesting that a part of the energy for increased milk production came from mobilization of body fat. Administration of bST effectively improved performance of cows under hot summer conditions whether evaporatively cooled or not.

Animals↗
PubMed1998

A new candidate site for a tumor suppressor gene involved in mouse thymic lymphomagenesis is located on the distal part of chromosome 4.

In a previous work we provided preliminary evidence of the existence of a putative tumor suppressor gene region on the distal part of chromosome 4 in gamma-radiation-induced T-cell lymphomas of (C57BL/6J x RF/J) F1 hybrid mice. This region, named as TLSR2 (Thymic Lymphoma Suppressor Region 2), was located centered at D4Mit54. A more detailed allelotype analysis in the mentioned tumors with new informative microsatellites on distal chromosome 4 region, as well as in thymic lymphomas induced with gamma-rays in F1 hybrids generated from reciprocal crosses between the strains C57BL/6J and BALB/cJ, and having in mind the new map position of D4Mit205b, allowed us to confirm the existence of TLSR2 and to define it more precisely as centered at D4Mit205b. In addition, we identified a new candidate region, named as TLSR3 (Thymic Lymphoma Suppressor Region 3), located between the Mom-1 locus and D4Mit68, as the site of another putative tumor suppressor region involved in thymic lymphomagenesis.

Animals↗
PubMed1998

An AC-repeat adjacent to mouse Cdkn2B allows the detection of specific allelic losses in the p15INK4b and p16INK4a tumor suppressor genes.

The cyclin-dependent kinase inhibitors p15INK4b and p16INK4a are involved in the development of a wide range of human and murine tumors. These tumor suppressor genes are inactivated by deletions frequently associated to point mutations in the coding regions or hypermethylation of their promoters. In this work, we describe a simple-sequence length polymorphism located in mouse Chromosome (Chr) 4, between the Cdkn2B (p15INK4b) and Cdkn2A (p16INK4a) genes, only 700 bp downstream of the Cdkn2B locus. This DNA region was analyzed in different inbred strains showing a variable AC-repetitive DNA sequence. We used this microsatellite to detect loss of heterozygosity of the Cdkn2A and Cdkn2B loci in gamma-irradiation-induced thymic lymphomas of C57BL/6J x RF/J F1 hybrids. Using this specific marker, we were able to locate additional allelic losses not detected by other microsatellites. Since the allelic losses can be detected by a simple PCR amplification, this AC-repetitive sequence is specially useful as a genetic marker for these Cdkn2 genes and specifically for the p15INK4b cell cycle inhibitor.

Alleles↗
PubMed1998

Release of mast cell mediators into the jejunum by cold pain stress in humans.

BACKGROUND & AIMS: The central nervous system regulates gut functions via complex interactions between the enteric nervous and immune systems. The aim of this study was to investigate whether mast cell mediators are released into the human jejunal lumen during stress. METHODS: A closed-segment perfusion technique was used to investigate jejunal release of tryptase, histamine, prostaglandin D2, and water flux in response to the cold pressor test in 8 healthy subjects and 9 patients with food allergy. In 6 food-allergic patients, jejunal biochemical responses to cold pain stress were compared with those induced by food intraluminal challenge. RESULTS: Cold pain stress elevated heart rate and blood pressure and increased luminal release of mast cell mediators and jejunal water secretion in both groups. Stress-induced release of tryptase and histamine, but not of prostaglandin D2 and water flux, was greater in food-allergic patients than in healthy volunteers. In food-allergic patients, jejunal biochemical responses induced by cold pain stress were similar to those induced by antigen challenge. CONCLUSIONS: These results show the ability of the central nervous system to modulate intestinal mast cell activity and suggest that mast cells have a role in stress-related gut dysfunction.

Adult↗
PubMed1998

Effects of silymarin in alcoholic patients with cirrhosis of the liver: results of a controlled, double-blind, randomized and multicenter trial.

BACKGROUND/AIMS: Silymarin has protective effects in different experimental conditions, but its efficacy in human liver cirrhosis has not been completely established. Therefore, this study was carried out to determine the effect of silymarin in alcoholics with liver cirrhosis with respect to survival and clinical and laboratory changes. METHODS: From February 1986 to June 1989, we enrolled 200 alcoholics with histologically or laparoscopically proven liver cirrhosis in a randomized, double-blind multicenter trial comparing 450 mg of silymarin (150 mg/ three times per day) with placebo. The primary outcome was time to death, and the secondary outcome was the progression of liver failure. Additional analyses were also performed in 75 patients in whom anti-hepatitis C virus antibodies were measured after completion of the trial. RESULTS: One hundred and three patients were assigned to receive silymarin and 97 to receive placebo. The two groups were well matched for demographic and baseline clinical and laboratory features. A 2-year study period was completed in 125 patients (57 receiving silymarin and 68 receiving placebo). Twenty-nine patients (15 receiving silymarin, and 14 receiving placebo) died during the trial. Survival was similar in patients receiving silymarin or placebo. The effect of silymarin on survival was not influenced by sex, the persistence of alcohol intake, the severity of liver dysfunction or by the presence of alcoholic hepatitis in the liver biopsy. Silymarin did not have any significant effect on the course of the disease. No relevant side-effects were observed in any group. CONCLUSIONS: The results of this study indicate that silymarin has no effect on survival and the clinical course in alcoholics with liver cirrhosis.

Adult↗
PubMed1998

Kinetic profile of carbamazepine in an adult Portuguese outpatient population.

OBJECTIVE: The aim of our work was to define the kinetic profile of carbamazepine (CBZ), in order to improve on dosing schedules through a Bayesian approach. METHOD: Carbamazepine dose/steady-state trough concentrations data pairs and associated information were collected retrospectively on a population of adult epileptic patients. RESULTS: Fifty patients (index population) with two or more available concentrations (total of 174 determinations) met our inclusion criteria. Patients were taking CBZ (200-1800 mg/day) in mono- or polytherapy regimens. The analysis assumed a one-compartmental model with first-order absorption and elimination. Due to the data source (only trough concentrations were measured as part of hospital routine), the volume of distribution was fixed at 1.19 l/kg. The final estimates for CL were: 0.075 +/- 0.027 (mono- and polytherapy), 0.069 +/- 0.020 (monotherapy), and 0.106 +/- 0.037 l/h/kg (polytherapy). In order to validate these results, we assessed their predictive capacity using 18 new patients (validation population), submitted to the same inclusion criteria and using Prediction-Error analysis. The results suggested a different CL value for our population compared to earlier published clearance values. The results also pointed to an increased metabolic rate associated with polytherapy. The prediction capacity of the optimization method derived from a Portuguese population made in an a priori evaluation indicated a low error (-0.04 microg/ml), close to the theoretical zero value. CONCLUSION: Our results provide specific data on CBZ disposition in a Portuguese population and given the wide variability in the literature values, our data may help improve dosing of CBZ in Portuguese patients.

Adult↗